CN1654632A - Biological agent prepared by Arthrobacter AS-1 strain and its application - Google Patents
Biological agent prepared by Arthrobacter AS-1 strain and its application Download PDFInfo
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- CN1654632A CN1654632A CN 200410079650 CN200410079650A CN1654632A CN 1654632 A CN1654632 A CN 1654632A CN 200410079650 CN200410079650 CN 200410079650 CN 200410079650 A CN200410079650 A CN 200410079650A CN 1654632 A CN1654632 A CN 1654632A
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Abstract
The present invention relates to biological preparation prepared with arthrobacter AS-1 strain and its application, and belongs to the field of microbial technology. The microbial strain is preserved in the preservation number of CGMCC No. 1281. The biological preparation is prepared through the process including the steps of: test seed culture, shaker culture, seeding tank culture and liquid fermentation in fermenting tank. The culture process has strain inoculating amount of 0.5-2 %, adopts the culture medium comprising beef paste, yeast extractum, peptone and glucose and of pH 6.8-7.2. The fermentation is controlled in 28-30 deg.c and pH 7.0-7.2, the fermentation time in the seeding tank is 16-20 hr, the fermentation time in the fermenting tank is 24-48 hr, and the biological preparation is obtained after the bacteria number in the fermentation liquid reaches 7.0 E8/ml. The present invention has the advantages of short fermentation period, saving in power, low cost, no pollution, etc.
Description
Technical field:
The present invention relates to a kind of biotechnological formulation and application thereof, the microorganism belonging to genus technical field with the preparation of Arthrobacter AS-1 bacterial strain.
Background technology:
A kind of preference that tobacco sucks as people, nicotine volatilize in combustion processes and make flue gas be alkalescence and generation irritating smell, and pungency has negative effect to quality of tobacco.The height of nicotine content plays considerable effect to the quality of tobacco product, and the height of nicotine content of tobacco leaves directly has influence on the quality of quality, but the nicotine content of dissimilar tobacco leaves all has a moderate index.Nicotine content increases, and quality of tobacco is had a negative impact, and Here it is, and high-quality tobacco leaf needs one of reason of chemical ingredients coordination.
Good flue-cured tobacco upper leaf plays leading role in modern blended type cigarette and low tar Virginian-type cigarette tobacco leaf formulation, cigarette fragrance and style thereof are had very big contribution, and is in very great demand in the international market.By contrast, those quality levels are not high, style is not outstanding or the upper tobacco leaf of unfavorable industrial cost efficiency, and tobacco industry and commerce is not all had magnetism forever.People see that also under the influence of WTO and WHO, the flue-cured tobacco upper leaf will have ample scope for one's abilities.The problem matter of utmost importance that upper tobacco leaf exists on operability is that nicotine content of tobacco leaves is too high.Therefore reduce nicotine content of tobacco leaves, help to improve upper tobacco leaf operability, to improve the market competitiveness of sucking security, improving domestic tobacco leaf significant.
Dissimilar tobaccos, same type tobacco different soils are different to the influence of tobacco leaf nicotine with cultivation technique levels such as fertilising, irrigation conditionss, and nicotine content also changes with the variation of blade inserted part, and progressively improve leaf position from bottom to top nicotine content.The approach that reduces nicotine at present is: 1. the envrionment conditions of adjusting tobacco growing, use effective integrated agriculture measure: as using nitrogen phosphorus fertilizer in right amount, increase the usage quantity of potash fertilizer, with lime and the suitable cake fertilizer that uses, regulate the pH value, water by cigarette strain growth-development law, transplant in good time, according to practical situation, pinch in good time, suitably stay leaf, use growth regulator, proper mature (ripe) is gathered, optimize modulation condition and utilize ageing and fermentation to wait and comprehensively reduce nicotine content, there is nornicotine effect instability between year in this method, operation link is too much, peasant household disperses shortcomings such as not easy to operate.2. pass through to improve filtration efficiency in cigarette, dilution flue gas etc. though these measures can be regulated nicotine in the flue gas arbitrarily, often do not have suitable selectivity, make the human consumer be difficult to accept.3. utilize ageing and fermentative degradation nicotine and utilize the existing report of microbiological deterioration storage phase tobacco leaf nicotine, reported that the microorganism that can reduce nicotine comprises: Alcaligenes paradoxus alcaligenes paradoxus, Arthrobacter globiformils Arthrobacter globiformis, Arthrobacter nicotianae tobacco Arthrobacter, Arthrobacter nicotinovorans bites the nicotine Arthrobacter, Arthrobacter oxydans oxidation Arthrobacter, Bac illus megaterium bacillus megaterium, Bacillus mycoides bacillus mycoides, the Bacilluspumilus bacillus pumilus, Bacillus subtills subtilis, Cellulomonas sp. cellulomonas cartae, Corynebacterium sp. Corynebacterium, Enterobacter cloacae enterobacter cloacae, Micrococcusnicotianae tobacco micrococci, the false pseudomonas bacillus of Pseudomonas sp., Pseudomonas nicotianae tobacco pseudomonas, the false pseudomonas bacillus of Pseudomonas putida stench, Deharyomyces nicotianae, the little gram sweat of Cunninghamella echinulata thorn spore mould, Pellicularia filamentosa silk nuclear film grass bacterium, the yeast yeast.But this measure be difficult to solve bacterial strain use age, use back moisture content in leaves content and improve problems such as causing tobacco mildew easily, never adopt in the factory.Along with the growing interest of people to objectionable constituent in the flue gas, fragrance can be the technical barrier that tobacco faces for many years for the human consumer accepts still when reducing nicotine.
Summary of the invention:
The objective of the invention is mechanism based on the microbial metabolites degrading nicotine, study and found a kind of tobacco leaf nicotine of can degrading, improve the Arthrobacter AS-1 bacterial strain of quality of tobacco, after adopting this bacterial strain through the cultivation of test tube kind, bacterial classification shaking table enlarged culturing, seeding tank, fermentor tank liquid fermenting, be prepared into and a kind ofly can reduce tobacco smoke alkaloid content but do not influence tobacco leaf aesthetic quality's biotechnological formulation.
Biotechnological formulation of the present invention prepares through liquid fermentation production method by producing bacterial strain.Wherein:
1, producing bacterial strain is Arthrobacter (Arthrobacter sp.) AS-1, and this bacterial strain has been deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center, and preserving number is CGMCC NO.1281, preservation day to be on December 28th, 2004; This bacterial strain forms oyster white bacterium colony, G on beef-protein medium
+, aerobic, do not form gemma, the catalase positive, decomposition of cellulose does not think that this bacterial strain belongs to genus arthrobacter.
Arthrobacter (Arthrobacter sp.) AS-1 is the bacterial strain that the inventor found in 2004.This bacterial strain is gathered the fresh normal tobacco sample from the Yunnan Binchuan, the conventional serial dilution of process is coated to separate on the NA substratum and is obtained, this bacterial strain is measured through the external nicotine of routine, and discovery can significantly reduce the content of nicotine, makes nicotine-containing substratum become blueness by white.
2, liquid fermenting production prepares the process of biotechnological formulation and is:
The preparation of microbial inoculum:
(1) test tube is cultivated: adopt the NA substratum, this bacterial classification inoculation on the test tube slant substratum, was cultivated 1-2 days for 28 ℃.
(2) shake-flask culture: adopt the NA liquid nutrient medium, be seeded in the Erlenmeyer flask of filling liquid substratum cultivating bacterium in the above-mentioned test tube, in 30 ℃ of shaking culture 1-2 of shaking table days.
The liquid fermenting production process comprises that the cultivation of test tube kind, bacterial classification shaking table enlarged culturing, seeding tank, fermentor tank liquid fermenting prepare biotechnological formulation; Wherein:
A. the test tube kind is cultivated and is: the thalline of Arthrobacter sp.AS-1 is inoculated on the test tube culture medium slant, culture medium prescription is to add extractum carnis 3g in 1000 ml distilled waters, yeast extract 1g, peptone 5g, glucose 10g, control pH6.8-7.2 cultivated 1-2 days down at 28-30 ℃, obtained the test tube kind;
B. bacterial classification shaking table enlarged culturing is: the inclined-plane seed is inoculated in the triangular flask liquid nutrient medium, culture medium prescription is to add extractum carnis 3g in 1000 ml distilled waters, yeast extract 1g, peptone 5g, glucose 10g, control pH6.8-7.2 cultivated 16-20 hour at 28-30 ℃, the shaking table of rotating speed 200-300rpm;
C. liquid seeds jar fermentation is: the bacterial classification that shaking table is enlarged is inoculated in the fermentor tank of differing capacities in the ratio of 0.5-2%, and culture medium prescription is the culture medium prescription of shaking table enlarged culturing, and culture temperature is 28-30 ℃, and incubation time is 16-20 hour;
D. liquid fermentation tank prepares biotechnological formulation and is: the bacterial classification that will enlarge step by step through the seeding tank fermentation is inoculated in the fermentor tank of differing capacities in the ratio of 1-3%, culture medium prescription is the culture medium prescription of bacterial classification shaking table enlarged culturing, culture temperature is 28-30 ℃, pH7.0-7.2, incubation time is 24-48 hour, and bacterial number reaches 7.0 * 10 in fermented liquid
8Individual/as during ml, to obtain biotechnological formulation.
The invention has the advantages that: 1. fermentation period is short, and save energy, production cost are low; 2. microorganism is from vega soil, and to plant and person poultry harmless, easily production, anti-storage are easy to industrialization development, and be workable; 3. can fall the alkali flavouring, improve the quality of tobacco, and the nornicotine effect not influence the tobacco leaf aesthetic quality.
Embodiment:
Embodiment 1:(is used for cured tobacco leaf)
Biotechnological formulation of the present invention by producing bacterial strain (Arthrobacter sp.) AS-1 bacterial strain, is pressed the liquid fermentation production method preparation.It is identical that concrete preparation process and condition and summary of the invention are described part.
The application of biotechnological formulation of the present invention: the tobacco leaf that maturation is good is gathered by position (in, top), enter before the barn with biotechnological formulation of the present invention by 5% amount (content of biotechnological formulation in diluent) evenly spray in tobacco leaf, top, compile bar then and go into stove, according to a conventional method baking.Tobacco leaf after the baking shows after measured, and biological system of the present invention reaches 34.68% to the nicotine degradation rate of upper tobacco leaf, and the nicotine degradation rate of middle part tobacco leaf is reached 36.86%.
Cured tobacco leaf aesthetic quality smoking result shows, adopt biotechnological formulation of the present invention to handle after, tobacco leaf strength obviously descends, perfume quantity obviously improves, and fragrance matter makes moderate progress, and pungency obviously weakens, assorted gas obviously alleviates (withered and burnt smell, the assorted gas of powder), and mouthfeel is better than contrast.
Embodiment 2:(is used for the burley tobaccos tobacco leaf)
Biotechnological formulation of the present invention by producing bacterial strain (Arthrobacter sp.) AS-1 bacterial strain, is pressed the liquid fermentation production method preparation.It is identical that concrete preparation process and condition and summary of the invention are described part.
The application of biotechnological formulation of the present invention: will cut the cigarette strain of keeping well and enter the room that dries in the air, and after one week of system of drying in the air, evenly spray at position, burley tobaccos upper, middle and lower the system of drying in the air according to a conventional method by 5% amount (content of biotechnological formulation in diluent) with biotechnological formulation of the present invention.Burley tobaccos after the system of drying in the air show after measured, and biotechnological formulation of the present invention reaches 46.56%, the nicotine degradation rate of middle part burley tobacco leaf is reached 51.82.% the nicotine degradation rate of top burley tobacco leaf, and the nicotine degradation rate of bottom burley tobacco leaf is reached 54.63%.
Burley tobaccos tobacco leaf aesthetic quality smoking result shows, adopt biotechnological formulation of the present invention to handle after, tobacco leaf strength obviously descends, perfume quantity obviously improves, fragrance matter makes moderate progress, pungency obviously weakens, sensory evaluating smoking's quality is better than the CK contrast.
Claims (2)
1. biotechnological formulation with Arthrobacter AS-1 bacterial strain preparation, press the liquid fermentation production method preparation by producing bacterial strain, the production bacterial strain that it is characterized in that this biotechnological formulation is Arthrobacter (Arthrobacter sp.) AS-1 bacterial strain, this bacterial strain has been deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center, and preserving number is CGMCC N0.1281; The process that liquid fermenting production prepares biotechnological formulation is:
(1). the liquid fermenting production process comprises that the cultivation of test tube kind, bacterial classification shaking table enlarged culturing, seeding tank fermentation, fermentor tank liquid fermenting prepare biotechnological formulation; Wherein:
A. the test tube kind is cultivated and is: the thalline of Arthrobacter sp.AS-1 is inoculated on the test tube culture medium slant, culture medium prescription is to add extractum carnis 3g in 1000 ml distilled waters, yeast extract 1g, peptone 5g, glucose 10g, control pH6.8-7.2 cultivated 1-2 days down at 28-30 ℃, obtained the test tube kind;
B. bacterial classification shaking table enlarged culturing is: slant strains is inoculated in the triangular flask liquid nutrient medium, culture medium prescription is to add extractum carnis 3g in 1000 ml distilled waters, yeast extract 1g, peptone 5g, glucose 10g, control pH6.8-7.2 cultivated 16-20 hour at 28-30 ℃, the shaking table of rotating speed 200-300rpm;
C. liquid seeds jar fermentation is: the bacterial classification that shaking table is enlarged is inoculated in the fermentor tank of differing capacities in the ratio of 0.5-2%, and culture medium prescription is the culture medium prescription of shaking table enlarged culturing, and culture temperature is 28-30 ℃, and incubation time is 16-20 hour;
D. liquid fermentation tank prepares biotechnological formulation and is: the bacterial classification that will enlarge step by step through the seeding tank fermentation is inoculated in the fermentor tank of differing capacities in the ratio of 1-3%, culture medium prescription is the culture medium prescription of bacterial classification shaking table enlarged culturing, culture temperature is 28-30 ℃, pH7.0-7.2, incubation time is 24-48 hour, and bacterial number reaches 7.0 * 10 in fermented liquid
8Individual/as during ml, to obtain biotechnological formulation.
2, the described biotechnological formulation with the preparation of Arthrobacter AS-1 bacterial strain of claim 1 is used for tobacco and falls the alkali flavouring, improves the quality of tobacco.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 200410079650 CN1264971C (en) | 2004-12-30 | 2004-12-30 | Biological agent prepared by Arthrobacter AS-1 strain and its application |
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| CN 200410079650 CN1264971C (en) | 2004-12-30 | 2004-12-30 | Biological agent prepared by Arthrobacter AS-1 strain and its application |
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| CN1654632A true CN1654632A (en) | 2005-08-17 |
| CN1264971C CN1264971C (en) | 2006-07-19 |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101485334B (en) * | 2009-02-27 | 2011-05-11 | 云南省烟草科学研究所 | Biological microbial formulation for increasing potassium content in tobacco leaf |
| CN102119784A (en) * | 2011-02-23 | 2011-07-13 | 红云红河烟草(集团)有限责任公司 | Spice extract prepared from waste tobacco leaves and preparation method and application thereof |
| CN102643772A (en) * | 2012-05-10 | 2012-08-22 | 广西中烟工业有限责任公司 | Microbe strain and application thereof |
| US8895472B2 (en) | 2009-05-12 | 2014-11-25 | Tobacco Research Institute Of Hubei Province | Agent for reducing nicotine and harmful components in tobacco leaves for use in field cultivation of tobacco |
| CN108903049A (en) * | 2018-09-14 | 2018-11-30 | 四川省食品发酵工业研究设计院 | A method of using biotechnology alcoholization fermentation " fresh and sweet fragrant " type cured tobacco leaf |
-
2004
- 2004-12-30 CN CN 200410079650 patent/CN1264971C/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101485334B (en) * | 2009-02-27 | 2011-05-11 | 云南省烟草科学研究所 | Biological microbial formulation for increasing potassium content in tobacco leaf |
| US8895472B2 (en) | 2009-05-12 | 2014-11-25 | Tobacco Research Institute Of Hubei Province | Agent for reducing nicotine and harmful components in tobacco leaves for use in field cultivation of tobacco |
| CN102119784A (en) * | 2011-02-23 | 2011-07-13 | 红云红河烟草(集团)有限责任公司 | Spice extract prepared from waste tobacco leaves and preparation method and application thereof |
| CN102119784B (en) * | 2011-02-23 | 2013-05-01 | 红云红河烟草(集团)有限责任公司 | Spice extract prepared from waste tobacco leaves and preparation method and application thereof |
| CN102643772A (en) * | 2012-05-10 | 2012-08-22 | 广西中烟工业有限责任公司 | Microbe strain and application thereof |
| CN102643772B (en) * | 2012-05-10 | 2014-03-12 | 广西中烟工业有限责任公司 | Microbe strain and application thereof |
| CN108903049A (en) * | 2018-09-14 | 2018-11-30 | 四川省食品发酵工业研究设计院 | A method of using biotechnology alcoholization fermentation " fresh and sweet fragrant " type cured tobacco leaf |
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| CN1264971C (en) | 2006-07-19 |
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Granted publication date: 20060719 Termination date: 20211230 |